RESUMO
Glutathione peroxidase 3 (GPx3) is the only extracellular selenoprotein (Sel) that enzymatically reduces H2O2 to H2O and O2. Two GPx3 (CqGPx3) cDNAs were characterized from crayfish Cherax quadricarinatus. The nerve cord CqGPx3a isoform encodes for a preprotein containing an N-terminal signal peptide of 32 amino acid residues, with the mature Sel region of 192 residues and a dispensable phosphorylation domain of 36 residues. In contrast, the pereiopods CqGPx3b codes for a precursor protein with 19 residues in the N-terminal signal peptide, then the mature 184 amino acid residues protein and finally a Pro-rich peptide of 42 residues. CqGPx3 are expressed in cerebral ganglia, pereiopods and nerve cord. CqGPx3a is expressed mainly in cerebral ganglia, antennulae and nerve cord, while CqGPx3b was detected mainly in pereiopods. CqGPx3a expression increases with high temperature and hypoxia; meanwhile, CqGPx3b is not affected. We report the presence and differential expression of GPx3 isoforms in crustacean tissues in normal conditions and under stress for high temperature and hypoxia. The two isoforms are tissue specific and condition specific, which could indicate an important role of CqGPx3a in the central nervous system and CqGPx3b in exposed tissues, both involved in different responses to environmental stressors.
Assuntos
Astacoidea , Selênio , Aminoácidos/genética , Animais , Astacoidea/genética , Astacoidea/metabolismo , Clonagem Molecular , DNA Complementar/genética , Peróxido de Hidrogênio/metabolismo , Hipóxia , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Sinais Direcionadores de Proteínas/genética , Selênio/metabolismo , TemperaturaRESUMO
This study was aimed at determining the role of the crustacean hyperglycemic hormone (CHH) in the physiological compensation to both saline and thermal stress, in the freshwater crayfish Cherax quadricarinatus. By determining the expression of the CHH gene in the eyestalk of juvenile crayfish, we found that maximal induction of CHH was induced at high salinity (10 g/L) and low temperature (20 °C). In order to investigate the role of CHH in the physiological compensation to such stressful conditions, recombinant CHH was supplied to stressed animals. CHH-injected crayfish showed increased hemolymphatic levels of glucose, in accordance with a significant utilization of glycogen reserves from the hepatopancreas. Furthermore, CHH administration allowed stressed animals to regulate hemolymphatic sodium and potassium at more constant levels than controls. Taken together, these results suggest a relevant role of CHH in increasing the energy available intended for processes involved in the physiological compensation of C. quadricarinatus to both saline and thermal stress.
Assuntos
Proteínas de Artrópodes/metabolismo , Astacoidea/metabolismo , Temperatura Baixa , Metabolismo Energético , Água Doce/química , Hormônios de Invertebrado/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Salinidade , Tolerância ao Sal , Estresse Fisiológico , Sequência de Aminoácidos , Animais , Proteínas de Artrópodes/administração & dosagem , Proteínas de Artrópodes/genética , Astacoidea/genética , Clonagem Molecular , Ecossistema , Metabolismo Energético/efeitos dos fármacos , Glucose/metabolismo , Glicogênio/metabolismo , Hemolinfa/metabolismo , Hepatopâncreas/metabolismo , Hormônios de Invertebrado/administração & dosagem , Hormônios de Invertebrado/genética , Dados de Sequência Molecular , Proteínas do Tecido Nervoso/administração & dosagem , Proteínas do Tecido Nervoso/genética , Potássio/metabolismo , Proteínas Recombinantes/administração & dosagem , Transdução de Sinais , Sódio/metabolismoRESUMO
Lectins participate in the immune mechanisms of crustaceans. They have been considered as humoral receptors for pathogen-associated molecular patterns; however, some reports suggest that lectins could regulate crustacean cellular functions. In the present study, we purified and characterized a serum lectin (CqL) from the hemolymph of Cherax quadricarinatus by affinity chromatography and determined its participation in the regulation of hemocytes' oxidative burst. CqL is a 290-kDa lectin in native form, constituted by 108, 80, and 29-kDa subunits. It is mainly composed of glycine, alanine, and a minor proportion of methionine and histidine. It showed no carbohydrates in its structure. CqL is composed of several isoforms, as determined by 2D-electrophoresis, and shows no homology with any crustacean protein as determined by Lc/Ms mass spectrometry. CqL agglutinated mainly rat and rabbit erythrocytes and showed a broad specificity for monosaccharides such as galactose, glucose, and sialic acid, as well as for glycoproteins, such as porcine stomach and bovine submaxillary mucin and fetuin. It is a Mn(2+)-dependent lectin. CqL recognized 8% of crayfish granular hemocytes and increased 4.2-fold the production of hemocytes' superoxide anion in vitro assays when compared with non-treated hemocytes. This effect showed the same specificity for carbohydrates as hemagglutination; moreover, superoxide dismutase and diphenyleneiodonium chloride were effective inhibitors of CqL oxidative-activation. The CqL homoreceptor is a 120-kDa glycoprotein identified in the hemocytes lysate. Our results suggest that CqL participates actively in the regulation of the generation of superoxide anions in hemocytes using NADPH-dependent mechanisms.
Assuntos
Astacoidea/química , Astacoidea/imunologia , Hemócitos/imunologia , Lectinas/análise , Testes de Aglutinação , Aminoácidos/análise , Animais , Western Blotting , Cromatografia de Afinidade , Eletroforese em Gel Bidimensional , Hemócitos/metabolismo , Hemolinfa/metabolismo , Imuno-Histoquímica , Lectinas/sangue , Lectinas/imunologia , Espectrometria de Massas , Fagocitose/imunologia , Espécies Reativas de Oxigênio/metabolismo , Explosão Respiratória/imunologia , Estatísticas não ParamétricasRESUMO
Determinamos el momento del desarrollo postembrionario en que se produce la diferenciación sexual primaria en la langosta Cherax quadricarinatus. Esta es evidenciada por la presencia de las gónadas y sus respectivos conductos. También determinamos la diferenciación sexual definida por la aparición de los caracteres sexualessecundarios. Se observaron 797 machos, 506 hembras y 456 individuos intersexos de 0.02 a 89.96 g (de criadero y laboratorio). Disecamos una submuestra de 106 machos, 69 hembras y 59 individuos intersexos para la caracterización macroscópica de la estructura gonadal. La diferenciación de los gonoporos se inicia aproximadamente a los 0.10 g, en sincronía con la diferenciación del sistema reproductor en machos, hembras e intersexos. La adquisición de la forma definitiva de ovario, oviducto, testículo y vaso deferente son posteriores. El appendix masculina iniciasu diferenciación a los 0.12-0.2 g y adquiere los rasgos característicos del appendix de los adultos a partir de 1-2 g.La diferenciación de la mancha roja (red patch) ocurre a partir de los 2.3 g.
We determined the earliest stage of postembryonic development at which primary and secondary sexual differentiations occur in the freshwater (red claw) crayfish Cherax quadricarinatus. For this purpose,797 males, 506 females and 456 intersex specimens within a weight range of 0.02-89.96 g were observed under stereoscopic microscope to determine the presence of the genital openings at the basis of the third (females) or fifth (males) pair of pereiopods. Animals presenting both pairs of genital openings were considered as intersex. A subsample of 106 males, 69 females and 59 intersex were dissected for the macroscopic characterization of gonad morphology. The development of the genital openings began approximately at 0.10 g, simultaneously with the differentiation of the reproductive system in females, males and intersex. Although the differentiation of the reproductive system started very early in the postembryonic development, the definitive form and colour of ovaries, oviducts, testes and vasa deferentia was acquired later. The differentiation of the appendix masculina began at 0.12-0.2 g and acquired the elongated shape of the adult at 1-2 g. The soft red patch characteristic of adult males started at 2.3 g of body weight in both chelipeds.
Assuntos
Animais , Astacoidea/fisiologia , Diferenciação Sexual/fisiologia , Transtornos do Desenvolvimento Sexual , Astacoidea/anatomia & histologia , Caracteres SexuaisRESUMO
Cherax quadricarinatus is a large freshwater crayfish species Parastacidae) native of north-west Queensland and the Northern Territory of Australia. The species typically exhibits a gonochoristic sexual system, although in cultured populations various types of intersex individuals have been described as functional males. In the present study, the macroscopic morphology and the gonadal histology of one type of intersex are described and discussed. All intersexes having both pairs of genital openings (female and male openings) and lacking both appendix masculinae and red patches were functional females with normal ovaries and oviducts. From a histological point of view, they did not differ from normal females having previtellogenic and/or vitellogenic ovaries according to size.
Cherax quadricarinatus, es un astácido dulceacuícola de gran tamaño de la (familia Parastacidae) originario delnoroeste de Queensland y del norte de Australia. Presenta un sistema sexual gonocórico, aunque en poblaciones decultivo se han descrito varios tipos de individuos intersexos como machos funcionales. En el presente estudio se describe y discute la morfología macroscópica y la histología gonadal de un tipo de intersexos. Todos los intersexos que presentan ambos pares de aberturas genitales (femeninas y masculinas) y carecen de ambos apéndices masculinos y de la mancha roja, fueron hembras funcionales con ovarios y oviductos normales. Desde el punto de vista histológico no difieren de las hembras normales, presentando ovarios previtelogénicos y/o vitelogénicos de acuerdo a su tamaño.
Assuntos
Animais , Astacoidea/anatomia & histologia , Gônadas/anatomia & histologia , Transtornos do Desenvolvimento Sexual , Astacoidea/fisiologia , Gônadas/ultraestrutura , Reprodução/fisiologiaRESUMO
Extracted venoms from two species of cubozoan, Chironex fleckeri (C. fleckeri)and Chiropsalmus sp., were injected into the freshwater crayfish Cherax quadricarinatus (C. quadricarinatus). By means of a Vascular Doppler, the cardiac parameters heart rate and contraction amplitude were recorded, with the theoretical total heart output calculated from these two recordings. Cherax quadricarinatus reflects prey items utilized by these two species in the wild and so comparisons of venom action on this animal have greater ecological relevance than those of previous mammalian models used. While the results of this study reflect previous research showing that the overall potency of C. fleckeri venom is greater than that of Chiropsalmus sp. venom, the action of the venoms on the heart muscle is found to be different. Chironex fleckeri,which has the most potent venom out of these two species, is actually less effective in reducing cardiac activity than Chiropsalmus sp., which may be a direct reflection of the variation in primary prey utilized by each species.
RESUMO
Extracted venoms from two species of cubozoan, Chironex fleckeri (C. fleckeri)and Chiropsalmus sp., were injected into the freshwater crayfish Cherax quadricarinatus (C. quadricarinatus). By means of a Vascular Doppler, the cardiac parameters heart rate and contraction amplitude were recorded, with the theoretical total heart output calculated from these two recordings. Cherax quadricarinatus reflects prey items utilized by these two species in the wild and so comparisons of venom action on this animal have greater ecological relevance than those of previous mammalian models used. While the results of this study reflect previous research showing that the overall potency of C. fleckeri venom is greater than that of Chiropsalmus sp. venom, the action of the venoms on the heart muscle is found to be different. Chironex fleckeri,which has the most potent venom out of these two species, is actually less effective in reducing cardiac activity than Chiropsalmus sp., which may be a direct reflection of the variation in primary prey utilized by each species.